WO2004103094A1

WO2004103094A1 - Use of a soluble lucerne extract in order to slow weight gain in mammals

Info

Publication number: WO2004103094A1
Application number: PCT/FR2004/001203
Authority: WO
Inventors: Didier Coulmier
Original assignee: Desialis
Priority date: 2003-05-16
Filing date: 2004-05-14
Publication date: 2004-12-02
Also published as: FR2854769B1; US20060182822A1; EP1641356A1; CA2526061A1; FR2854769A1

Abstract

The invention relates to the use of a soluble Lucerne extract obtained by pressing fresh Lucerne, containing at least 1 wt. % saponines in relation to the weight of the extract, for the production of a food preparation which can limit weight gain in mammals, partially in the case of a hyperlipemic diet, said preparation being adapted for addition to the food rations of the mammal in doses of said extract amounting to 0.8g to 3g per day and per kilogram of mammal body weight .

Description

USE OF A SOLUBLE EXTRACT FROM LUZERNE TO LIMIT WEIGHT GAIN IN MAMMALS

The invention relates to the use of a soluble extract of alfalfa in the field of human and animal nutrition, and in particular of mammals.

The diets of Western populations lead to many dysfunctions, some of which are obesity and high blood cholesterol levels, which are the cause of many diseases.

The synthetic substances developed by the pharmaceutical industry to fight against these metabolic dysfunctions have, in a certain number of cases, many side effects, some of which can be lethal. Weight gain like an individual's cholesterolemia is partly linked to their eating habits and in particular to the share of lipids ingested.

The blocking of part of the lipids during the digestive process leading to non-absorption at the intestinal level, makes it possible to reduce the energy supply of the ration in the form of lipids, and to decrease the entry of the precursors of cholesterol in l 'organization.

It is known to those skilled in the art that saponins, which are especially present in many plants, have cholesterol-lowering effects. Thus, according to the article "Alfafa saponins and their Implication in

Animal Nutrition ”, J. Agric. Food Chem. 1998, 46, 131-140, it is known that the administration of saponins to animals such as chicks, adult roosters makes it possible to decrease the level of cholesterol in the blood plasma. Similarly, it is possible to prevent hypercholesterolemia in the Alps, induced by the ingestion of cholesterol, by the incorporation of saponins in the diet. However, nothing is said in this article about an effect of saponins on pea intake. In addition, nothing is said about the daily dosage of saponins necessary to obtain a significant effect on the level of blood cholesterol. Furthermore, French patent application FR 2 794 616 describes in particular a plant product obtained from alfalfa, concentrated in saponin or in sapogenin, and free from the unwanted accompanying carnavanine because it is toxic. French patent application FR 2 794 616 also describes the use of this plant product in a process for the production of a nutritional supplement having cholesterol lowering properties with regard to cholesterol present or introduced into the intestine. In this patent application, it is indicated that the daily absorption by a human being of this plant product containing 1.5 to 40% by weight of saponins or sapogenins, at a rate of 50 to 1000 mg per day, and in especially 200 to 400 mg per day, allows plasma cholesterol to be kept at a normal level, even when the person who takes it eats the products known to be loaded with cholesterol. However, nothing is said in this application concerning an effect of this product on the limitation of weight gain induced by such a diet. In addition, the dosage indicated does not take into account the weight of the individual.

However, the Applicant has discovered that the administration to rats, within the framework of a lipid-lowering diet, of an extract of soluble alfalfa containing at least 1% of saponin, at a rate of 0.8 to 3 g per day and per kilogram of animal live weight, allowed to limit weight gain The Applicant has also discovered that it made it possible to lower total cholesterol and triglyceride levels in the blood, while keeping VLDL cholesterol levels at constant values despite the lipid-lowering diet.

By VLDL cholesterol level (also called "bad cholesterol") is meant within the meaning of the present invention, the cholesterol level contained in very low density lipoproteins (very low density lipoprotein). Thus, the subject of the present invention is therefore the use of a soluble alfalfa extract obtained by pressing fresh alfalfa, and containing at least 1% by weight of saponins relative to the weight of the extract, to limit the uptake of weight in mammals, in particular within the framework of a hyperlipemic regimen, consisting in adding to the food ration of the mammal 0.8 to 3 g, preferably 0.9 g to 2.73 g, and better 0.91 g per day and per kilogram of live weight of the mammal of said extract.

Preferably, the soluble alfalfa extract used according to the invention is in the form of a powder having a moisture content of between 3 and 10% by weight, and comprises, by weight relative to the weight of dry powder :

- 15 to 25% crude protein,

- 10 to 30% of mineral matter,

- 0 to 2% of raw cellulose, - 10 to 25% of sugars, and

- 5 to 20% organic acids.

The mineral materials are conventionally based on potassium, calcium, magnesium, phosphorus, zinc and iron.

The sugars present in the soluble alfalfa extract used according to the invention are generally essentially represented by glucose and fructose, and to a lesser extent by galactose, stachyose and verbascose.

Advantageously, the soluble alfalfa extract comprises, by weight relative to the weight of dry powder, 5 to 12% of malic acid. The soluble alfalfa extract used according to the invention can be obtained by the process described in patent FR 2 294 647. It generally corresponds to an alfalfa juice obtained by pressing, freed of its precipitable proteins, then concentrated. The alfalfa of origin preferably does not undergo pre-haulage, which makes it possible to preserve a maximum of metabolic compounds present in the plant (vitamins, growth factors, intermediate metaboiites for example) at the time of harvest.

In particular, the soluble alfalfa extract can be obtained by concentrating and drying a serum obtained from fresh alfalfa according to a process comprising the following steps:

(a ') cold pressing of fresh alfalfa, which provides a green juice and a cake;

(b ') treatment of the green juice by alkalization at a pH in the region of 8.5, for example with ammonia; (c ') flocculation of green juice proteins by heat treatment, which provides a protein precipitate and a soluble extract (also called serum);

(d) centrifuging the soluble extract-precipitated protein mixture until the soluble alfalfa extract (or serum) is obtained;

(e ') drying the soluble extract of alfalfa by spraying or “spray drying”, which provides a dry extract in powdery form of ocher color.

The soluble alfalfa extract is preferably incorporated into a food preparation (liquid or solid) before being administered orally, in addition to the food ration.

Thus, the soluble alfalfa extract as defined above allows the manufacture of a food preparation capable of limiting weight gain in mammals, said preparation being suitable for adding to the food ration of the mammal of 0.8 g 3 g per day and per kilogram of live weight of mammal of said extract.

The use according to the present invention of an alfalfa extract as defined above also makes it possible, concomitantly with limiting weight gain, to decrease the cholesterol and total triglyceride levels in the blood, and to keep the rate constant VLDL cholesterol in the blood, even on a lipid-lowering diet.

The following example illustrates the invention in more detail, but not limitation.

EXAMPLE

Evaluation of the effects of an orally administered alfalfa extract on weight gain and lipemia in adult male Wistar rats.

In this example, we studied the effects of consumption for 30 days of an alfalfa extract (or alfalfa serum) containing 1.1% by weight of saponins relative to the weight of the extract, on weight gain and lipemia in adult male Wistar rats.

Three groups of 12 adult male rats received a hyperlipid diet for 45 days in order to induce a slight hyperlipemia (triglycerides and cholesterol). Two of these groups were administered, the alfalfa serum from 15 ^th to 45 days. The third group of rats, not treated with alfalfa serum, serves as a control.

Blood samples are taken during the induction of lipemia on days 0.7 and 15 and during treatment with alfalfa serum on days 30 and 45, to determine the lipid status of the animals.

1. Topics

Thirty-six adult male Wistar / AF EOPS rats (Charles River breeding center / Iffa-Credo, 69-St-Germain sur l'ArbresIe, France) aged 16 weeks.

The rats are marked and distributed in groups of two in polycarbonate type F cages (43 x 28x 15 cm, U.A.R., 91 - Epinay- sur-orge, France). The animals are kept in an air-conditioned animal house, at a temperature of 22-24 ° C and subjected to a reverse light-dark cycle of 12 hours (light from 8 p.m. to 8 a.m.).

After familiarization with laboratory conditions for 7 days, and following 15 days of lipid-lowering treatment, the rats are divided into 3 groups of homogeneous treatments in relation to their lipid status (cholesterol and triglycerides): - a control group (consisting of 12 rats ), and

- two groups "Luzerne Serum" (each also made up of 12 rats). The rats of the three groups are all handled in the same way and under the same conditions. In order to avoid possible interference between the various products, the two rats in the same cage receive the same product.

2. Tested product

Soluble alfalfa extract supplied in the form of dry powder by the company ALFALIS.

The characteristics of this product are presented below in Table 1. TABLE 1

3. Experimental procedure

Drinking water and the hyperlipid diet prepared specifically to increase the lipemia of animals are presented ad libitum to animals.

This diet consists of standard food (M20 food, Dietex, France) supplemented with lipids in the form of refined pork fat (Saindoux at 21.4%), cholesterol (1%, Acros Organics, Noisy-ie-Grand, France) , as well as in sucrose (3.8%). The prepared diet is reconstituted in the form of pellets by adding 20% water.

The rats of the two groups "Luzerne Serum" receive daily, from 15 * ™ to 45 ^th day, 0.91 g and 2.73 g / kg of body weight of the extract "Luzerne Serum", containing 10 and 30 mg saponins / kg.

The daily treatment of control rats consists of a solution of 0.54 g / kg of sucrose equivalent to the amount of carbohydrate contained in the extract of "Alfalfa Serum" at the most concentrated dose.

On days 0, 7, 15, 30 and 45, the rats of three groups are fasted 12 hours before the blood samples.

The administration protocol for the alfalfa extracts is indicated below in Table 2. TABLE 2

4 - Administration product extract "Serum Alfalfa" is suspended in an aqueous solution and administered orally daily from the ^15th to ^45th day.

5 - Variables studied For the application of the effect of alfalfa extract on the lipemia of male WISTAR rats, the following biological variables were determined in the serum: the total cholesterol level, the triglyceride level, and the VLDL cholesterol.

The weights of the animals are recorded every three or four days in order to adapt the doses of the treatments.

The rats' food intake is evaluated once a week.

6. Results

• Weight change

The weight change (in g) of the 3 groups of rats is represented in terms of average weight (in g), from day 1 to day 43, in FIG. 1, and corresponds to the results presented in Table 3. FIG. 1 and Table 3 show the following facts: - during the first 15 days of treatment with the lipid-lowering diet, the weights of the rats of the three groups change in an equivalent manner; - from day 18, and until the end of the treatment, the weight of the rats of the Luzerne 2 group remains significantly lower than that of the rats of the control group;

- from day 25 and until the end of the treatment, the weight of the rats of the Luzerne 1 group remains significantly lower than that of the rats of the control group. Treatment with alfalfa extract helps limit weight gain as a result of a lipid-lowering diet. The effect of limiting weight gain is more marked when the alfalfa extract is administered at 2.73 g / kg (Alfalfa 2), than at 0.91 g / kg (Alfalfa 1).

TABLE 3 Weight change (g) (average + ESM)

• Evolution of food intake

The dietary evolution from day 1 to day 43 of the 3 groups of rats is represented in terms of average food intake relative to body weight (in%) in FIG. 2, and corresponds to the results presented in table 4.

Figure 2 and Table 4 show that on days 1, 8, 16, 22, 29, 36 and 43, no significant heterogeneity was observed in the food intake of the rats of the 3 groups. The rats treated with the alfalfa extract gain significantly less weight than the control rats while the rats of the three groups consume the same amount of food relative to their respective weights.

TABLE 4 Evolution of food intake as a function of body weight (%)

(mean ± ESM)

• Evolution of total cholesterol level

The evolution of the total cholesterol level in the rats of the 3 groups, from day 1 to day 43, is represented in terms of average total cholesterol level in the blood (in g / l) in FIG. 3 and corresponds to the results presented in table 5.

These results show the following facts:

- during the 45 days of treatment with the hyperlipemic system, the total cholesterol levels remain constant in the rats of the Vehicle and Lucerne 2 groups;

- in rats of the Luzerne 1 group, the total cholesterol level decreases significantly from D30 to D45;

Treatment with alfalfa extract for 30 days at 0.91 g / kg lowers the total cholesterol level. TABLE 5

Evolution of total cholesterol level (g / 1)

(mean ± ESM)

• Evolution of the triglyceride level

The evolution from day 1 to day 43 of the triglyceride level is presented in terms of average blood level in g / l in FIG. 4, and corresponds to the results presented in table 6. These results show the following facts:

- during the first 15 days of treatment with the hyperlipemic regimen, the triglyceride level increases significantly in rats of the three treatment groups;

- from D15 to D30, it decreased significantly in the rats of the Vehicle and Alfalfa 1 groups;

- from D30 to D45, it decreased significantly in the rats of the Luzerne 1 and Luzerne 2 groups; - on the last day of the treatments (D45), the triglyceride level is lower in the rats of the Luzerne 1 and Luzerne 2 groups than those of the control group.

Treatment with alfalfa extract for 30 days can significantly decrease the triglyceride level when administered at 0.91 g / kg and tends to decrease it when administered at 2.73 g / kg. TABLE 6 Evolution of the triglyceride level (g / l) (mean ± ESM)

• Evolution in VLDL cholesterol level

The evolution of the VLDL cholesterol level, from day 1 to day 43, of the groups of rats is represented in terms of average blood level in FIG. 5, and corresponds to the results presented in table 7. The results show that:

- during the first 15 days of treatment with the lipid-lowering diet, the VLDL cholesterol level remains constant in the rats of the Vehicle and Luzerne 1 groups, but increases significantly in the rats of the Luzerne 2 group. This increase is due to significant individual variability within this group independent of the administration of alfalfa which only started from day 15;

- from D15 to D45 the VLDL cholesterol level tends to increase in the control rats despite a decreasing tendency between D30 and D45;

- on day 45 the VLDL cholesterol level in the control rats is significantly higher than that of the rats in the group

Alfalfa 1 and tends to be higher than that of rats in the Luzerne 2 group;

- from D15 to D45 the VLDL cholesterol level tends to decrease in the rats of the Luzerne 2 group, and decreases significantly from D30 to D45 to regain a normal basal value; - during the treatment period, the VLDL cholesterol level remained constant in the rats of the Luzerne 1 group.

Treatment with alfalfa extract for 30 days keeps VLDL cholesterol at constant values despite the lipid-lowering diet.

Treatment of rats with alfalfa extract seems more effective at a dose of 0.91 g / kg / day than when administered at a dose of 2.73 g / kg / day.

TABLE 7 Evolution of VLDL cholesterol level (g / l) (mean ± ESM)

The effects on weight gain and lipemia of the consumption of alfalfa extract for 30 days in adult male Wistar rats are therefore the following:

- limitation of weight gain resulting from a hyperlipemic diet; - at a dose of 0.91 g / kg, decrease in the total cholesterol level;

- significant decrease in the triglyceride level when the alfalfa extract is administered at 0.91 g / kg, and tendency to decrease when it is administered at 2.73 g / kg;

- maintaining the VLDL cholesterol level at constant values despite the lipid-lowering diet; the alfalfa extract seems more effective at the dose of 0.91 g / kg / d than at the dose of 2.73 g / kg / d.

Claims

1. Use of a soluble alfalfa extract obtained by pressing fresh alfalfa and containing at least 1% by weight of saponins relative to the weight of the extract, for the manufacture of a food preparation capable of limiting weight gain in mammals, in particular within the framework of a hyperlipemic system, said preparation being suitable for adding to the food ration of the mammal from 0.8 g to 3 g per day and per kilogram of live weight of mammal of said extract.

2. Use according to claim 1, characterized in that the food preparation is suitable for adding to the food ration from 0.9 g to 2.73 g, and preferably 2.73 g per day and per kilogram of live weight of mammal of said extract.

3. Use according to claim 1 or 2, characterized in that the soluble alfalfa extract is in the form of a powder having a moisture content of between 3 and 10% by weight, said extract comprising, by weight relative to the weight of dry powder:

- 15 to 25% crude protein,

- 10 to 30% of mineral matter, - 0 to 2% of raw cellulose,

- 10 to 25% of sugars, and

- 5 to 20% organic acids.

4. Use according to claim 3, characterized in that the soluble alfalfa extract comprises, by weight relative to the weight of dry powder, 5 to 12% of malic acid.

5. Use according to any one of the preceding claims, characterized in that the soluble alfalfa extract is obtained by concentration and drying of a serum obtained from fresh alfalfa according to a process comprising the following stages: (a ') pressing cold fresh alfalfa, which provides a green juice and an oil cake;

(b ') treatment of the green juice by alkalization at a pH in the region of 8.5, for example with ammonia;

(c ') flocculation of the proteins of the green juice by heat treatment, which provides a protein precipitate and a soluble extract of alfalfa; (d ') centrifuging the soluble extract-precipitated protein mixture until the soluble alfalfa extract is obtained;

(E ') drying the serum by atomization, which provides a dry extract in powdery form of ocher color.

6. Use according to any one of the preceding claims, characterized in that said extract is incorporated into the food preparation before being administered orally, in addition to the food ration.

7. Use according to any one of the preceding claims, characterized in that, concomitantly with the limitation of weight gain, said extract also makes it possible to reduce the level of total cholesterol in the blood.

8. Use according to any one of claims 1 to 6, characterized in that, concomitantly with the limitation of weight gain, said extract also makes it possible to keep constant the level of VLDL cholesterol in the blood.

9. Use according to any one of claims 1 to 6, characterized in that, concomitantly with the limitation of weight gain, said extract also makes it possible to reduce the level of triglycerides in the blood.